Details

Haruyo Asahina,Chief of the Narcotic Section of the National Hygienic Laboratory;Toyohiko Kawatani,Director of the Kasukabe Experiment Station of Medicinal Plants, Attached to the National Hygienie Laboratory;Miss Masako Ono,Narcotic Section, National Hygienie Laboratory;Sanaenosuke Fujita, Kasukabe Experiment Station of Medicinal Plants, Tokyo, Japan

Introduction

Since the promulgation of the Japanese Opium Act in 1954, which allows the cultivation of the opium poppy under licence, we have grown the opium poppy and other poppies for study and experiments, and the opiums or juices therefrom have been analysed.

Papaver setigerum DC. and its hybrid with P. somniferum L. have been grown. The main alkaloids in opiums from these different poppies were estimated and the percentages compared.

On the opium poppy varieties and other poppies, we tested for the presence or absence of morphine by our method of paper chromatography. The chromosome numbers of various species were counted. Further, for varieties or strains of P. somniferum, the morphine content was determined for the purpose of the plant breeding, chiefly by our method of paper chromatography and spectrophotometry.

The content of "porphyroxine-meconidine ", a red-turning alkaloid when heated in diluted mineral acid, of Japanese opium and of opium from successive lancings of P. somniferum was measured spectrophoto-metrically.

The morphine content of commercial opiums produced by the Japanese cultivators in the crop seasons of 1953/54 and 1954/55 was assayed by us by the method of the Japanese Pharmacopoeia, and this information is summarized in the paper.

Our paper is divided into the following sections:

Alkaloidal analysis of the opium from Papaver setigerum DC.

A natural hybrid of Papaversetigerum DC. and P. Somniferum L.

Examination of poppies of various species for the presence of morphine.

Examination of poppy varieties of Papaver somniferum L. for opium of the highest morphine yield.

The porphyroxine content of Japanese opium and of opium from successive lancings.

The morphine content of Japanese opium.

Acknowledgements

It is a pleasure to acknowledge our indebtedness to Dr. Charles G. Farmilo, Organic Chemistry and Narcotic Section, Food and Drug Directorate, Canada, for the seed of P. setigerurn DC. and to the chemists of the laboratory of the Division of Narcotic Drugs, United Nations, for their invaluable and continuous help and guidance.

We wish to express our thanks to S. Mizumachi and Y. Shiuchi, National Hygienic Laboratory, for their assistance in making many of the analyses during this investigation. Thanks are also due to T. Ohno, Kasukabe Experiment Station, for his aid in cytogenetical studies.

The presence of morphine in Papaver setigerum DC., previously suspected, was confirmed by Farmilo et al.[1] in 1953. Morphine was detected in the pod, bud, and leafy parts of this poppy plant.

The Japanese Opium Act (Act No. 71) promulgated in 1954, prohibits the cultivation of the opium poppy, including P.setigerum, except under licence.

For actual production of opium only P. somniferum has been cultivated, and there is no history of the cultivation of P.setigerum even experimentally, in Japan. No estimation of the alkaloids in opium from this poppy plant has been reported prior to the writing of the present paper. The capsules of P.setigerum are small and dehiscent, and this species can scarcely have any commercial use. Besides our desire to clarify the legal point of its inclusion in the Opium Act, it was studied for scientific reasons and as a possible parent in hybridization.

This investigation was undertaken to grow and identify botanically P. setigerum, and to determine quantitatively the content of alkaloids in the opium from this poppy plant.

With this in mind, the following two experiments, experiment I and experiment II, were made in successive years. The seeds were sown in October 1954 and 1955. The flowering continued almost throughout May of the following year.

There were plants with violet flowers and also plants with mauve flowers in the strain. In experiment I both kinds were used without separating them, whereas in experiment II only plants with violet flowers were used.

Three or four incisiom, each covering a quarter of the capsule, were made at intervals of one day, till no more latex was yielded. The first incision began ten days after the petals had fallen. In experiment I it began at different dates in the sequence of flowering, irrespective of the strains, whereas in experiment II all sets of incisions made began on the same date. Each incision consisted of four longitudinal slashes.

The capsules were incised in the evening and the latex was gathered just after incision (" Kiritori" collection) and the following morning (" Oigaki" collection). The opiums thus obtained were mixed, as is the usual practice in collecting commercial opium from P.somniferum.

Details of the Collection of Opium from Papaver setigerum DC.

Experiment

Seeds sown

Strains used

Capsules incised

Opium per capsule (air.dried)(Mg)

Period o/ incision

Plant height(cm)

I

20.X.1954

Violet and mauve-flowered

90

15.56

16-31.V.1955

65

II

25.X.1955

Violet-flowered

130

15.88

18-24.V.1956

67

The presence of alkaloids, at the proper Rf values for morphine, codeine, and other opium alkaloids in the opium from P. setigerum was readily demonstrated by using paper chromatography [2]

Opium was rubbed thoroughly with dilute hydrochloric acid, and the solution containing the alkaloidal salts was centrifuged. The supernatant solution was used for the paper chromatography.

The paper chromatography was carried out as follows:

Solvent: n-butanol 50, 28 % NH4OH 9, distilled water 15 parts by volume. In the preparation of the solvent, the constituents were mixed, the mixture allowed to stand for some time, and the top layer of the mixture used for chromatography.

Spot-developing reagent: Potassium iodoplatinate prepared according to Munier: 1 cc of 10% platinic chloride and 25 cc of 4% potassium iodide were mixed and the resulting solution made up to 50 cc with H 20.

The following spots were obtained. The Rf values and colouration of the spot are given.

Chromatography of Opium from Papaver setigerum DC.

Opium

R/ values

Experiment I

0.73

0.87'

0.92

Experiment II

0.72

0.88

0.93

(Opium tincture)

0.71

0.87

0.93 for comparison; P. somniferum)

Colouration

Dark blue

Violet

Violet

The characteristic dark blue colouration of the morphine spot on a filter paper, when treated with potassium iodoplatinate solution, and occurring at the proper Rf value for morphine, near 0.71 with this solvent, was easily recognized.

The other alkaloidal spots were coloured violet. The Rf value of the codeine spot was 0.87 and the spot of Rf 0.93 corresponded to other by-alkaloids. Thebaine, papaverine, and narcotine were inseparable from each other by this solvent.

Opium tincture was used for comparison to fix the position of the morphine spot more precisely under the particular conditions.

The morphine content of the opium from P. setigerum was then determined by paper chromatography and spectrophotometry. [3] [4]

The contents of codeine, thebaine, papaverine, and narcotine were also assayed by our unified method.[5]

The results, as determined on the air-dried opium, are given below:

Alkaloidal Analyses of Opium from Papaver setigerum DC.

Opium

Morphine

Codeine

Thebaine

Papaverine

Narcotine

Experiment I

5.1

0.9

2.1

1.9

0.1

Experiment II

7.3

0.8

1.6

2.6

0.1

The values were quite similar for the two crop seasons. Perhaps this is due to a dose control of growing and harvesting methods and use of a particular strain of the species, but it is naturally not known whether this species may vary much in the alkaloids present in the opium or not. We hope later to examine one or more other strains of P. setigerum.

For comparison, the alkaloidal analyses of a few opium samples from the United Nations Opium Distribution Centre assayed by our unified method are given below. These relate, of course, to opium from P. somniferum.

Comparative Alkaloidal Analyses of Opium from Papaver somniferum L.

Origin of opium

Morphine

Codeine

Thebaine

Papaverine

Narcotine

Turkey export U.N.15

13.5

1.7

0.8

0.8

4.9

India excise U.N.36

11.6

4.2

1.8

0.4

6.3

Iran Fars U.N.47

12.8

4.0

3.5

1.5

7.1

The results show that opium from P. setigerum is comparatively low in morphine and codeine, but high in thebaine and papaverine, and very low in narcotine. While morphine is still the principal alkaloid, narcotine is not the second one, but either thebaine or papaverine, and in general the picture of alkaloidal composition is almost opposite to that of somniferum opium.

The chromosome number was confirmed to be n = 22 in the haploid; 2n = 44 in the diploid (fig. 1 and 2).

Vesselovskaya mentioned in 1933 that P. setigerum, although distinct, could be crossed readily with P. somniferum, indicating their close genetic relationship, and that the crosses were perfectly normal fertile hybrids with viable seed.[6] However, we do not know of either a chemical or a detailed botanical investigation of such a hybrid before our own.

It was observed in 1955 that the violet- and mauve-flowered strains of P. setigerum were almost pure phenotypically, that is, the violet- or mauve-flowered strain consisted almost exclusively of plants with violet or mauve flowers respectively.

An F 1 hybrid was found to grow in the violet-flowered strain in 1956. The chromosome number in the root-tip cell was 2n= 33, which was the stun of the gametic chromosome numbers of P. setigerum n = 22, and P. somniferum n = 11 (fig. 3). At the first meiotic metaphase in the pollen mother cell (PMC), 11 bivalent and 11 univalent chromosomes were observed (fig. 4). The chromosome behaviour in meiosis of PMCs of the F 1 hybrid was of the type of Pilosella.

Apparently the female parent is the violet-flowered strain of P. setigerum, whereas the male parent is considered to be P. somniferum, "Ikkanshu" variety, a strain commonly cultivated for actual production of opium in Japan. In 1955, no species with a chromosome number 2n = 22 or its multiple other than P. somniferum and P. setigerum was grown by us, and "Ikkanshu" was cultivated in the adjoining field to the female parent at our experiment station.

External characteristics as well as habits of the hybrid were observed in detail. It was matroclinous. The leaves, stems, flowers and capsules were like those of the female parent but were larger in size.

Seven capsules of the hybrid were incised in the usual way, as described in section 1, and 0.24 g of opium was obtained.

The analytical values for alkaloides assayed by our unified method[7] on the opiums from the hybrid and its parents are listed as follows. The parents were grown again in the same year as the hybrid(1956).

Comparative Alkaloidal Analyses of opium from the F 1 Hybrid and its Parents

(Percentage)

Opium

Morphine

Codeine

Thebaine

Papaverine

Narcotine

P. setigerum (?)

7.3

0.8

1.6

2.6

0.1

F 1 Hybrid

13.2

3.6

1.5

2.6

0.2

P. somniferum (?)

"Ikkanshu"

16.0

3.7

1.0

0.9

1.1

By comparison with the result for opium from the female parent, P.setigerum, the opium of the hybrid showed a fairly considerable increase in morphine content (almost doubled) and a large increase in codeine content (four times as great). Thebaine and papaverine, already high, were not changed.

The narcotine content increased from 01% to 0.2% by our determination, but thus remained remarkably low.

As compared with the opium from the male parent. P. somniferum, the opium of the hybrid was almost equal in morphine and codenine, very low in narcotine, but quite high in thebaine and papaverine.

It is very interesting to observe that in the production of some alkaloids the hybrid became almost equal to the female parent, but in others it remained like like the female parent, whether the production was high, as with thebaine and papaverine, or low, as with narcotine.

Summary :

A natural hybrid of P. setigerum DC (?) and P. somniferum L. (?) was found. Connecting the external characteristics, it was matroclinuos.

The chromosome number was 2n=33. At the first meiotic metaphase in PMCs ,11 bivalent and 11 univalent chromosomes were observed.

The chromosome behaviour in meiosis of PMCs was of the type of Plosella.

Morphine was detected in the opium from the hybrid. The content was 13.2%.

The opium of the hybrid was high in thebaine and papaverine, but very low in narcotine. In morphine and codeine production, the hybrid was superior to the female parent and was almost equal to the male parent; but in production of other alkaloids, thebaine, papaverine, and narcotine, it corresponded to the female parent.

During 1955 and 1956 a considerable number of varieties and strains of Papaver somniferum, P. orientale, and P. rhoeas, together with 29 other species grown at the Kasukabe Experiment Station from seeds sent by foreign botanical gardens, were examined for the presence of morphine by our method of paper chromatography. The method has been given in a preceding section of this paper and was previously published in a United Nations document.[8]

The results are set out in Table II. In nearly all cases the latex was used for the test. In no poppy but P. somniferum and P. setigerum was any clear evidence of morphine found. In five other cases (Nos. 36 to 39, and No. 45) there was a spot at about the right Rf value, but the colouration was different from that of a morphine spot.

Examination was made by the ordinary paraffin method to determine or confirm the chromosome numbers of 31 of these species of Papaver. The results are set out in Table III -This table also shows in which cases previous determinations are known to us. It will be noted that only P.somniferum and P. setigerum were found to have chromosome numbers which are multiples of 11.

TABLE II

Detection of morphine in poppies by paper chromatography Rf value and colouration of the alkaloidal spots

L: Latex

B: Blue

C: Capsule

DB: Dark blue

S: Straw

BG :Blue green

V: Violet

DV: Dark violet

Italics: Morphine spot

No.

Poppy

Rf relative to 100 and Colouration

Origin

Papaver

1

somniferum .

L

71DB

88V

94V

Japan, perhaps originally

69DB

86V

from North China

69DB

87V

92V

71DB

86V

91V

2

Somniferum

L

72DB

95V

Saitama, Japan

73DB

95V

70DB

92V

3

Rhoeas

L

none

Tokyo, Japan

4

Rhoeas

L

91V

Tokyo, Japan

93V

5

orientale "Mahoney"

L

15DB

96V

Tokyo, Japan

6

orientale bracteatum

L

18DB

36V

94V

Tokyo, Japan

7

orientale "Queen Alexandra"

L

18DB

95V

Tokyo, Japan

8

orientale "Feltham"

L

17BG

25V

94V

Tokyo, Japan

9

orientale "Beauty of Livermere"

L

18BG

25V

96V

Tokyo, Japan

10

orientale "Salmon Queen"

L

95V

Tokyo, Japan

11

orientale "Apricot Queen" .

L

18DB

26V

92V

Tokyo, Japan

12

orientale "Princess Victoria Louise"

L

19BG

27V

96V

Tokyo, Japan

16BG

23V

96V

17BG

24V

96V

17BG

25V

92V

13

orientale "Rembrandt"

L

19BG

20DB

28V

91V

Tokyo, Japan

14

orientale "Pink Beauty" .

L

18BG

27V

91V

Tokyo, Japan

15

orientale "Mrs. Perry"

L

19DB

29V

93V

Tokyo, Japan

16

orientale "Columbia (Olympia fl. pl.)"

L

19DB

94V

Tokyo, Japan

17

orientale "Perry's White"

L

18BG

26DB

28V

93V

Tokyo, Japan

18BG

26V

89V

18

somniferum

L

77DB

94V

Tokyo, Japan

19

somniferum .

L

78DB

96V

Tokyo, Japan

20

somniferum

L

72DB

94V

Tokyo, Japan

70DB

92V

21

somniferum

L

71DB

88V

93V

Tokyo, Japan

22

setigerum

L

74DB

89V

93V

Saitama, Japan

73DB

87V

92V

67DB

88V

94V

23

somniferum

L

72DB

94V

Tokyo, Japan

24

somniferum

L

76DB

95V

Tokyo, Japan

25

somniferum

L

75DB

91V

96V

Tokyo, Japan

26

somniferum

L

77DB

96V

Tokyo, Japan

27

somniferum .

L

75DB

91V

95V

Tokyo, Japan

28

somniferum .

L

70DB

92V

Tokyo, Japan

29

somniferum .

S

73DB

Tokyo, Japan

30

somniferum

S

70DB

84V

Japan, perhaps originally from North·China

C

73DB

88V

31

rhoeas

C

none

Tokyo, Japan

32

orientale

L

20BG

28DB

92V

Kassel, Germany

22BG

30DB

95V

20BG

29 DB

94V

33

argemone

L

none

Louvain Belgium

34

rhoeas

L

93V

Rome, Italy

35

pyrenaicum

L

90V

Hamburg, Germany

36

polare

L

17V

21DB

75DV

95V

Hamburg, Germany

37

atlanticum

L

16V

21DB

74DV

94V

Vienna, Austria

38

rubriaurantiacum

L

16V

20DB

72DV

92V

Helsinki, Finland

TABLE II (continued)

Detection of morphine in poppies by paper chromatography

Rf value and colouration of the alkaloidal spots

No.

Poppy

Rf relative to 100 and Colouration

Origin

Papaver

39

rupifragum var. atlanticum

L

18V

21DB

41DB

72DV

96V

Helsinki, Finland

17V

23DB

69DV

91V

40

rhoeas

L

92V

Helsinki, Finland

41

alpinum

L

none

Uppsala, Sweden

42

apulum

L

none

Uppsala, Sweden

43

glaucum

L

86V

Uppsala, Sweden

44

lecoqii

L

95V

Uppsala, Sweden

45

pilosum

L

17V

22DB

70DV

93V

Uppsala, Sweden

46

rhoeas "Shirley Wilks"

L

89V

Wageningen, Netherlands

47

lateritium

L

18V

23DB

40V

89V

Berlin-Dahlem, Germany

48

rubro-aurantiacum

L

18V

24DB

41V

91V

Basle, Switzerland

49

pavoninum

L

none

London, United Kingdom

50

umbrosum

L

94V

Klagenfurt, Austria

51

aurantiacum

L

16V

19DB

37DB

90V

Munich, Germany

52

mairei

L

21DB

88V

Munich, Germany

53

monanthum

L

93V

Munich, Germany

54

persicum

L

17V

20DB

94V

Munich, Germany

55

oreophilum

L

92V

Munich, Germany

56

argemone

L

93V

Cologne, Germany

57

bracteatum

L

22DB

30DB

94V

Cologne, Germany

58

nudicaule

L

none

Cologne, Germany

59

californicum

L

92V

Dublin, Eire

60

hybridum

L

none

Dublin, Eire

61

pyrenaicum

L

92V

Dublin, Eire

62

triniaefolium

L

91V

Dublin, Eire

63

lateritium

L

16V

19DB

90V

Dublin, Eire

64

nudicaule

L

12V

86V

Heidelberg, Germany

65

monanthum

L

12DV

14DB

3lB

67V

86V

Sofia, Bulgaria

66

dubium

L

3V

Toulouse, France

67

collinum

L

2V

Nantes, France

68

hybridum L

L

none

Uppsala, Sweden

69

rhoeas var. latifolium

L

11DB

91V

London, United Kingdom

70

schinzianum

L

14DB

91V

Braumchweig, Germany

71

rupifragum

L

13DB

88V

Warsaw, Poland

72

orientale

L

3V

12BG

18B

88V

Graz, Austria

TABLE III

Chromosome numbers of Papaver species (determinations of T. Kawatani and T. Ohno)

Species

Chromosome number 2n

Previous determinations a b 2n

species

Chromosomenumber2n

Previous determinations a b 2n

Papaver

Papaver

alpinum

14

14 (Ljungdahl, 1922) (Sugiura, 1936,

hybridum

14

14 (Ljungdahl, 1922)

1940) (Faberge, 1944)

(Sugiura, 1937, 1940)

apulum

14

12 (Sugiura, 1936)

lateritium

14

14 (Ljungdahl, 1922) Yasui, 1941)

argemone

42

12 (Beale, 1939);

lecoqii

14

42 (Sugiura, 1936, 1940)

mairei

14

atlanticum

14

14 (Ljungdahl, 1922)

monanthum

14

12, 14 (Sugiura, 1940)

nudicaule

14

14 (Ljungdahl, (1922)(Fabere, 1944)

aurantiacum

14

28 (Horn, 1938)

bracteatum

42

14 (Yasui, 1936)

oreophilum

14

14 (Sugiura, 1936)

californicum

14

orientale

42

42 (Yasui, 1921) (Ljungdahl, 1922)

collinum

28

28 (Snoad, 1952) 42 (Yamazzaki, 1936)

glaucum

14

14 (Sugiura, 1931, 1944)

pavoninum

12

12 (Sugiura, 1931, 1936)

(Kuzmina, 1935)

persicum

14

14 (Ljungdahl, 1922)

TABLE III (continued)

Chromosome numbers of Papaver species (determinations of T. Kawatani and T. Ohno)

For 98 varieties or strains of Papaver somniferum, using in most cases seeds from foreign botanical gardens to obtain the plants, the morphine content of the opium was determined and compared with that of opium poppy plants of ordinary Japanese origin.

When sufficient opium could be collected from plants of one strain, the method of assay of the Japanese Pharmacopoeia VI was used. When the amount of opium was smaller, this method was modified to use only 1 gramme of opium. When less than 1 gramme was the most that could be collected, our method of paper chromatography and spectrophotometry was used).[9]

The results are given in Table IV.

The poppies were sown in October 1954 and 1955 and grown under uniform cultural conditions at the Kasukabe Experiment Station (numbers 1 to 26, 58 to 98) and the Wakayama Experiment Station (numbers 27 to 57). The method of incision was the usual practice described in section 1.

It is remarkable that the ten highest results for morphine percentage are on opium of poppies of which the seeds came from certain quite northern botanical gardens, and they are above 20%; but the quantity of opium was low.

Variety No.

Morphinepercentage

Seed origin

25

30.0

Copenhagen, Denmark

64

27.5

Tabor, Czechoslovakia

11

24.9

Cologne, Germany

22

24.0

Montreal, Canada

19

23.6

Cologne, Germany

20

23.0

Munich, Germany

67

22.1

Nantes, France

90

21.2

Li?, Belgium

91

21.0

Seattle, United States of America

63

20.7

Hamburg, Germany

In these cases the determinations were all made by our paper chromatographic method. This method possibly yields somewhat higher results than the precipitation method of the Japanese Pharmacopoeia, on the same opium.

However, it is to be noted that the highest results by the modified J.P. VI method were the following:

Variety No.

Morphinepercentage

Seed origin

36

20.04

Copenhagen, Denmark

47

19.82

Cologne, Germany

It is very interesting to observe that two samples of variety monstrosum, No. 25 and 67, gave very high results; 30% and 22.1%.

Contrary to the high morphine percentage in some opiums of foreign seeds, the ordinary Japanese opium "Ikkanshu" gave medium results.

However, opium weight per capsule was also calculated. The poppies of Japanese origin, "Ikkanshu", produced a large quantity of opium. As can be seen in Table IV, the poppies which gave more opium than 150 mg per capsule were all of the "Ikkanshu" strain. Therefore the Japanese strain "Ikkanshu" is superior in productivity of opium to all other poppies that we have grown in Japan from foreign seeds; but in these trials so far we have used seeds from botanical gardens and not seeds from poppies used for commercial opium production in other countries.

The poppies producing opium of exceptionally high morphine percentage, such as Danish and Czech poppies, produced a very small quantity of opium per capsule; but such poppies have great interest for trials as parent stock for commercial production, and our paper chromatographic method is a very convenient means for selecting plants of high productivity.

The content of "porphyroxine-meconidine" of opium from successive lancings of the same poppy capsules was determined.

The method used was based on the one described by L. Fuchs and W. Ullrich.10 Some modifications were introduced.

Experimental

About 0.15 g of opium was accurately weighed. In a small mortar, 0.1 g of calcium hydroxide and 2.5 cc of water, precisely measured, were added and the mixture rubbed to a smooth paste. After addition of 12.5 cc of water, precisely measured, the mixture was stirred for 20 minutes.

It was then filtered. 10 cc of the filtrate, precisely measured and brought into the extraction apparatus (fig. 5), was extracted continuously with peroxide-free ether for about 4 hours until a negative test for "porphyroxine-meconidine" was obtained with the return ether.

The ether was then evaporated on the steam-bath. To the residue, 0.5 cc of dilute HC1 (1 part by volume of conc. HC1 to 99 of water) was added, and the solution was heated in a boiling water-bath. After 5 minutes the colour was fully developed. The red solution was rinsed in to make exactly 40 cc or 20 cc in a volumetric flask.

The absorbance of the red-coloured solution was measured by a Beckman 13 spectrophotometer at 511 mµ, using a 1 cm cell.

The value for 10 cc of solution obtained from 0.1 g opium was calculated.

Opium

( b) From Papaver somniferum L.-" Ikkanshu" cultivated during 1954/55 at the Wakyama Experiment Station attached to the National Hygienic Laboratory.

By "Kiritori" method.-" Kiritori" is a method of collecting the latex from poppy capsules. Its meaning is cutting (Kiri) and taking from (tori), that is, to incise the unripe capsule and immediately take the latex away from the poppy capsule.

By" Oigaki" method.-" Oigaki" is another method. It means an additional (Oi) collection (gaki), that is, to collect the latex (" Asabuki ") which blows out (buki) early the following morning (Asa) from the same incision made in the "Kiritori" method.

( d) From P. somniferum cultivated during 1954/55 by a drug company (Nippon Shinyaku) in Kyoto.

Calculated values for the absorbance of the coloured solution of 10 cc, obtained from 0.1 g of opium, measured with Beckman B spectrophotometer at 511 mµ.

Lancing

(a)

(b)

(c)

(d)

(e)

I

1. 524

2.175

2.129

1.157

0.755

II

1.915

3.167

2.267

0.380

1.115

III

2.487

2.407

2.145

IV

2.228

2.376

2.973

Volume in which measurement carried out (cc.)

40

40

40

20

20

Later experiments were made on opium front poppies grown from foreign seed obtained from botanical gardens with the following results:

Opium sample

Absorbance lancing

Country where seeds obtained

I

II.

f

0.728

1.550

Rome, Italy

g

0.123

0. 589

Amsterdam, Netherlands

h

0.594

1.448

Braunschweig, Germany

i

0.702

1.152

Turin, Italy

Calculated by the equivalent worked out by Farmilo and Kennett,11 the "Lovibond values" of the above readings should be the following (multiplication of the "1%" absorbance in a 1-cm cell by 30.2):

Calculated Lovibond values

Lancings

Opium sample

Seed origin

I

II

III

IV

a

46.0

57.8

75.1

67.3

Japan

b

65.7

95.2

72.7

71.7

Japan

c

64.3

68.5

64.8

89.8

Pakistan

d

34.9

11.5

Pakistan

e

22.8

33.7

India

f

22.0

46.8

Italy

g

3.7

17.8

Netherlands

h

17.9

43.7

Germany

i

21.2

34.8

Italy

In Table V are given the results of comparative colorimetric determinations of "porphyroxine-meconidine" in Japanese opiums, in comparison with opiums of five other countries (China, India, Iran, Korea, Turkey). Twelve Japanese samples of recent production are included, with one sample of the former legal production, and two samples (Nos. 402 and 435) from poppies grown experimentally in Japan. Only in the case of one of the latter samples is the value as low as for the opiums of China, Iran, Korea, or Turkey (in the samples examined).

In our earliest determinations (in 1954) comparisons were made with a photoelectric colorimeter, using filter S.53. These have been converted to calculated" Lovibond values ", for ready, comparison with values given by the UN Secretariat, by the factor obtained by Fuchs & Ullrich,12 equivalent to multiplication of the "1%" absorbance by 39.5. In the second part of the table are determinations made with the Beckman B spectrophotometer. For these the "Lovibond values" have been calculated using the equivalent found by Farmilo and Kennett, already mentioned.

MAP II. Villages of Opium Poppy Cultivation

Black spot shows the village in which the opium poppy was actually cultivated under licence during 1954/55.

Nagano

Wakayama

1.

Ooka, Sarashina

1.

Tasugawa, Arita

2.

Murakami, Sarashina

2.

Yuasa, Arita

3.

Sakagi, Hanishina

3.

Hiro, Arita

4.

Tanaka, Chiisagata

4.

Yura, Hidaka

5.

Ena, Hidaka

Aichl

Hiroshima

1.

Ono, Saeki

1.

Akabane, Atsumi

2.

Itsukaichi, Saeki

1.

Tawara, Atsumi

3.

Kurose, Kamo

4.

Suigano, Mitsugi

Osaka

5.

Fuchu

1.

Toyokawa, Mishima

Okayama

2.

Fukui, Mishima

1.

Sakuto, Aida

3.

Takatsuki

4.

Ibaragi

Hyogo

5.

Suita

1.

Anji, Shiso

Conclusions

The content of porphyroxine in Japanese opium is nearly always high, and sometimes extremely high. This is very characteristic.

The changes in the porphyroxime content with repeated lancings do not seem to be entirely regular, but in eight cases out of nine the content in opium of the second lancing was greater than in opium of the first lancing.

Method: All except 402, 435 by our K/15 method 13 402 and 435 by a modification of the Eder & Wäckerlin method 14

Lab. No.

Morphine %

Origin (in Japan)

401

13.99

Kambe, Atsumi, Aichi

402

10.75

Ichijoji, Sakyo, Kyoto

403

10.75

Fukui, Mishima, Osaka

404

14.24

Fukui, Mishima, Osaka

405

10.29

Ibaragi, Osaka

406

11.97

Ibaragi, Osaka

407

11.53

Takatsuki, Osaka

408

15.61

Minamihiro, Arita, Wakayama

409

15.37

Minamihiro, Arita, Wakayama

Lab. No.

Morphine %

Origin (in Japan)

410

15.64

Minamihiro, Arita, Wakayama

411

12.07

Tsugi, Arita, Wakayama

412

15.73

Minamihiro, Arita, Wakayama

413

11.80

Tsugi, Arita, Wakayama

414

13.87

Minamihiro, Arita, Wakayama

415

15.91

Minamihiro, Arita, Wakayama

416

17.60

Minamihiro, Arita, Wakayama

417

15.49

Tsugi, Arita, Wakayama

418

15.95

Hiro, Arita, Wakayama

419

12.55

Tsugi, Arita, Wakayama

420

11.64

Tsugi, Arita, Wakayama

421

15.23

Minamihiro, Arita, Wakayama

422

13.66

Minamihiro, Arita, Wakayama

423

14.08

Hiro, Arita, Wakayama

424

14.60

Hiro, Arita, Wakayama

425

13.30

Hiro, Arita, Wakayama

426

13.51

Hiro, Arita, Wakayama

427

14.25

Hiro, Arita, Wakayama

428

14.55

Hiro, Arita, Wakayama

429

15.18

Hiro, Arita, Wakayama

430

15.29

Minamihiro, Arita, Wakayama

431

15.08

Minamihiro, Arita, Wakayama

432

13.94

Minamihiro, Arita, Wakayama

433

10.62

Tsugi, Arita, Wakayama

434

11.96

Tsugi, Arita, Wakayama

435

14.12

Yada, Hidaka, Wakayama

In Table VI the laboratory analyses are given for Japanese opiums of the crop season of 1953/54, and in Table VII analyses are Summarized for the crop season of 1954/55.

As may be seen approximately (or calculated) from the latter table, the average content is only between 11% and 12 %, although opium of a much higher morphine percentage can be obtained by collecting the first lancings separately.

These determinations were made by the method of the Japanese Pharmacopoeia VI.

TABLE VII

Classification of Japanese opium samples according to morphine percentages, crop season 1954/55

M. A. Vesselovskaya. "The Poppy - Its classification and its importance as an oleiferous crop " - Supplement 56 to the Bulletin of Applied Botany, of Genetics and Plant Breeding. The Lenin Academy of Agricultural Sciences in the USSR, Leningrad; 1933 (in Russian and English).

Haruyo Asahina, "An assay procedure for opium based on that of the Japanese Pharmacopoeia, and morphine percentages obtained on samples from various countries ", United Nations document ST/SOA/SER. K/15, 11 March 1953. This procedure was adopted as official for the Japanese Pharmacopoeia as from 15 March 1955.